Measurement of albumin permeability across endothelial monolayers in vitro

J. A. Cooper, P. J. Del Vecchio, F. L. Minnear, K. E. Burhop, W. M. Selig, J. G. Garcia, A. B. Malik

Research output: Contribution to journalArticle

89 Scopus citations

Abstract

We have developed an experimental system to measure the permeability of the cultured endothelial monolayer. The luminal-to-abluminal flux of 125I-albumin across cultured pulmonary endothelium was expressed as a clearance rate equal to the permeability-surface area product. After clearance rate measurement for a 30-min base-line period, a test agent was added to the luminal side, and the clearance rate was remeasured during a 30-min experimental period. In control studies the base-line clearance rate was 0.343 ± 0.017 μl/min. After correction for the diffusional resistances of the filter and unstirred layers, the calculated permeability of the endothelial monolayer was 1.2 x 10-5 cm/s. When culture medium was the test agent, the experimental clearance rate was unchanged from the base-line value. After addition of 4 mM oleic acid to the luminal chamber, the clearance rate was 0.528 ± 0.017 μl/min compared with a base-line value of 0.330 ± 0.008 μl/min (P < 0.005). This method allows the calculation of endothelial permeability with correction for unstirred layers and the use of each monolayer as its own control.

Original languageEnglish (US)
Pages (from-to)1076-1083
Number of pages8
JournalJournal of Applied Physiology
Volume62
Issue number3
DOIs
StatePublished - 1987

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

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    Cooper, J. A., Del Vecchio, P. J., Minnear, F. L., Burhop, K. E., Selig, W. M., Garcia, J. G., & Malik, A. B. (1987). Measurement of albumin permeability across endothelial monolayers in vitro. Journal of Applied Physiology, 62(3), 1076-1083. https://doi.org/10.1152/jappl.1987.62.3.1076